The objective of this study was to investigate the flow and absorption of amino acids (AA) in steers fed diets supplemented with different protein sources. Twenty-one Holstein steers averaging eight months of age and 187 kg of body weight were used. Animals were fitted with abomasal caanulas and fed diets with a forage to concentrate ratio of 40:60. The experimental diets contained Rhodes grass hay, corn, molasses, minerals, and one of the following protein sources: cottonseed meal, yeast, or urea. Animals were assigned to treatments in a completely randomized design with three protein sources and seven replicates per treatment. Each experimental period lasted 17 days with 10 days for diet adaptation and seven days for sample collection. Samples of abomasal digesta were collected every 28 h while those of feces twice a day during the sampling period. The abomasal flow of AA (g/d) and the AA composition of abomasal and fecal samples were significantly affected by protein source. However, no protein source effect was observed for AA digestibility. The abomasal availability and intestinal absorption of AA were lower than expected probably because animals were feed restricted. It can be concluded that AA availability and composition were affected by the different protein sources fed to steers in this trial.